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Abstract

We present spatial light interference tomography (SLIT), a label-free method for 3D imaging
of transparent structures such as live cells. SLIT uses the principle of interferometric
imaging with broadband fields and combines the optical gating due to the micron-scale coherence
length with that of the high numerical aperture objective lens. Measuring the phase shift map
associated with the object as it is translated through focus provides full information about
the 3D distribution associated with the refractive index. Using a reconstruction algorithm
based on the Born approximation, we show that the sample structure may be recovered via a 3D,
complex field deconvolution. We illustrate the method with reconstructed tomographic refractive
index distributions of microspheres, photonic crystals, and unstained living cells.

Figures (6)

Fig. 1 Visualization of 3D sectioning of SLIM. (a) Sectioning effect of SLIM with
coherence gating. (b) An x-z cut through a live neuron; the bottom of the image
corresponds to the glass surface. The soma and nucleolus (arrow) are clearly visible.
(c-d) Images of the same neuron at the depths indicated by the dash lines in
(b). Scale bar for (b-d): 10 μm.

Fig. 6 Tomography capability. (a)-(b) Refractive index distribution
through a live neuron at position z = 0.4 μm (a) and 6.0 μm
(b). The soma and nucleolus (arrow) are clearly visible. Scale bars, 10
μm. (c) 3D rendering of the same cell. The field of view is 100
μm × 75 μm × 14 μm and NA = 0.75. (d)
confocal microscopy of a stained neuron with same field of view and NA = 1.2. Neurons were
labeled with anti-polysialic acid IgG #735. The 3D rendering in (c) and
(d) was done by ImageJ 3D viewer.